Nitric oxide secretion assay by murine macrophages


Nitric oxide (NO) plays a key role in the inflammatory response by activating several signaling pathways that exacerbate the deleterious effects of oxidative stress.

The quantification of nitric oxide (NO) is therefore a relevant biological target in the search for anti-inflammatory agents.


Nitrite accumulation is considered as an indicator of NO production in the culture medium.

NO is physiologically unstable and is rapidly oxidized to nitrite and nitrate. Nitrate is then reduced to nitrite by the action of nitrate reductase.

It is thus possible to indirectly quantify the production of NO via the determination of nitrite using the Griess reaction.

[Archer S.1993]


Format96 wells
Cellular modelRAW 264.7 (ATCC TIB-71)
Reference moleculeQuercetol ; L-NMMA
Incubation24H at 37°C under 5% CO
RevelationGriess reaction with absorbance measurement at 544 nm

Notes :

  • Allow 50 µL at 10 mM or 3 mg of sample to test 5 concentrations (3 points/concentration

  • Cell viability is assessed in parallel

Recommendation :

  • Ensure solubility of sample in a solvent compatible with cell culture

Inhibition of nitrite secretion in the presence of L-NMMA and Quercetol